PCR Kleen Spin Columns

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Overview

PCR Product and DNA Fragment Purification Guide

Bio-Rad offers a variety of pre-PCR and post-PCR sample preparation and cleanup columns and prepacked columns; many are also suitable for DNA fragment purification.

  Freeze 'N Squeeze DNA
Gel Extraction Spin Columns
Micro Bio-Spin and
Bio-Spin Columns
PCR Kleen
Spin Columns
Application Recovery of DNA fragments (50 bp–23 kb) purified from TAE- or TBE-buffered agarose gels Dye terminator and label cleanup from sequencing reactions; buffer exchange; desalting Removal of PCR products >200 bp from other reaction mixture components
Starting Material ≤700 mg gel slice 10–100 µl of sequencing reaction mixture 25–100 µl of PCR or other enzyme reaction
Format Spin-column extraction of frozen-thawed gel slices Prepacked size exclusion spin column Prepacked size exclusion spin column
Recovery 4–23 kb: >80% recovery;
0.5–4 kb: >70% recovery;
50–500 bp: >50% recovery
>95% of linear DNA >22 bases or bp; 99% retention of unincorporated dye terminators 50–80%
Time Required <10 min <10 min <4 min
Rapid Removal of Primer-Dimers

Effective, rapid removal of primer-dimers from PCR reactions. Two PCR reaction mixtures were purified with PCR Kleen spin columns. Lane 1, a mixture of 2 µg of a 660 bp DNA fragment with 100 ng of a 25-mer primer-dimer; lane 3, a mixture of 2 µg of a 660 bp DNA fragment with 500 ng of a 25-mer primer; lane 2, primer-dimer purification; lane 4, primer purification; M, DNA markers. Reaction products and purified samples (10 µl each) were analyzed on a 1.5% agarose gel.

Column Selection Guide
  Bio-Spin 6 Micro Bio-Spin 6 Bio-Spin 30 Micro Bio-Spin 30 PCR Kleen
Packed support Special grade
Bio-Gel P-6 gel
Special grade
Bio-Gel P-6 gel
Special grade
Bio-Gel P-30 gel
Special grade
Bio-Gel P-30 gel
Special grade
Size exclusion gel
Equilibration buffer 10 mM Tris, pH 7.4 or SSC buffer* 10 mM Tris, pH 7.4 or SSC buffer* 10 mM Tris, pH 7.4 or SSC buffer* 10 mM Tris, pH 7.4 or SSC buffer* 10 mM Tris, 1 mM EDTA, pH 7.0
Applications Desalting of oligonucleotides >20 bases - - -
Labeling reactions:
removal of unincorporated nucleotides >20 bases or bp from DNA
- - -
Removal of primers and primer-dimers from PCR products >200 bp - - - -
Buffer exchange
(restriction fragments, PCR products, enzyme reactions, sequencing templates.)
- -
DNA sequencing
reaction mixture cleanup**
- - -
Riboprobe cleanup*** - - - -
Desalting of antibody, enzyme, and protein solutions - -
Purification of proteins of moleular weight >6,000 - - -
Purification of proteins of molecular weight >40,000 - - -
Bed Volume 1.1 ml 0.7 ml 1.1 ml 0.7 ml 0.6 ml
Retention and recovery 90% recovery of 20 bases or bp, 99% retention of salts 90% recovery of 20 bases or bp, 99% retention of salts 95% recovery of 22 bases or bp, 98% retention of ddNTPs 95% recovery of 22 bases or bp, 98% retention of ddNTPs 85% recovery of ≥700bp, 95% retention of primers and primer-dimers
Molecular weight exclusion limit, globular proteins 6,000 6,000 40,000 40,000 8,000,000
Sample volume 50–100 µl 10–75 µl 50–100 µl 10–75 µl 25–100 µl
Centrifuge type Swinging bucket Microcentrifuge Swinging bucket Microcentrifuge Microcentrifuge
Autoclavability Yes Yes Yes Yes Yes

* 150 mM NaCl, 17.5 mM sodium citrate, pH 7.0
** In Tris buffer.
*** In RNase-free Tris buffer.

PCR Kleen columns are prepacked spin columns for purifying PCR products and other DNA molecules >200 bp directly from reaction mixtures. A simple 4 minute spin effectively removes salts, nucleotides, enzymes, primers, and primer-dimers. Purified DNA fragments are eluted into a collection tube and are immediately available for secondary PCR, subcloning, restriction digests, ligations, sequencing, and other enzymatic manipulations.

Method Size exclusion chromatography in spin column
Applications Removal of nucleic acids <32 bases and other reaction mixture components from PCR products >200 bp
Capacity 25–100 µl
Equllibration buffer 10 mm Tris, 1mM EDTA, pH 7.4
Sample preparation time <4 min
Yield 50–80% recovery
PCR Kleen Spin Columns

732-6300
Pkg of 25, prepacked spin column, for purification of PCR products and other DNA fragments >200 bp directly from reaction mixtures

List Price:   $101.00
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Number Description Options
2950 Comparison of PCR Kleen Spin Columns to Traditional Methods for Purification of PCR Products Prior to Sequencing, Rev A Click to download
2311 PCR Kleen Spin Columns Product Information Sheet, Rev D Click to download
4006142 Instruction Manual, Quantum Prep PCR Kleen Spin Columns, Biotechnology Explorer, Rev B Click to download